糖基化终产物对巨噬细胞MMP-9表达与活性的影响及其机制的初步研究

发布时间：2018-03-17 04:17

本文选题：糖基化终产物　切入点：糖尿病　出处：《中国医科大学》2006年硕士论文　论文类型：学位论文

【摘要】：前言在高血糖的条件下,许多生物大分子会发生非酶糖基化反应,产生糖基化终产物(Advanced Glycation End Products,AGEs),大量蓄积于体内并沉积于血管壁。近年大量研究证明,糖基化终产物在糖尿病性动脉粥样硬化的发病中起重要作用。基质金属蛋白酶参与动脉粥样硬化病变的进展,尤其是与斑块破裂导致的临床事件有密切关系。然而,糖基化终产物与基质金属蛋白酶的相关研究较少。本实验旨在通过观察糖基化终产物作用后的巨噬细胞的形态变化并测定MMP-9活性、表达改变,来研究糖基化终末产物对巨噬细胞活化及其与斑块破裂的关系;并根据他汀类药物的可阻断类异戊二烯途径的特性,结合类异戊二烯中间产物的逆转效应的观察,用以确证类异戊二烯通路是否参与了AGE对巨噬细胞基质金属蛋白酶的诱导过程。方法体外培养小鼠腹腔巨噬细胞,分别以不同浓度BSA-AGE(0、50、100、200、400mg/L)作用48h;以相同浓度BSA-AGE(400mg/L)作用不同时间(0、12、24、36、48h);在BSA—AGE(400mg/L)培养条件下,不同浓度的辛伐他汀(5μmol/L,50μmol/L)作用48h;以及BSA—AGE(400mg/L)与辛伐他汀(50μmol/L)培养条件下,GGPP(10μmol/L)作用48h。提取培养液上清,采用明胶酶谱法测定MMP-9活性;通过Western blot法检测MMP-9表达。图像光密度扫描后,统计学分析。
[Abstract]:Foreword. Under the condition of hyperglycemia, many biomolecules will undergo non-enzymatic glycosylation reactions, resulting in advanced Glycation End products ages, which are accumulated in the body and deposited in the vascular wall. Glycation end products play an important role in the pathogenesis of diabetic atherosclerosis. Matrix metalloproteinases are involved in the progression of atherosclerosis, especially in clinical events caused by plaque rupture. There are few studies on the relationship between the end products of glycosylation and matrix metalloproteinases. The aim of this study was to investigate the relationship between the activation of macrophages and plaque rupture by observing the morphologic changes of macrophages after glycosylation end products and the changes of MMP-9 activity and expression. According to the characteristics of statins blocking isoprene pathway and the reverse effect of intermediate product of isoprene, To confirm whether isoprene pathway is involved in the induction of matrix metalloproteinases by AGE. Method. Murine peritoneal macrophages were cultured in vitro and exposed to 400 mg / L BSA-AGEO at different concentrations for 48 h; 400 mg / L BSA-AGEO at the same concentration for 48 h; and 400 mg / L BSA-AGEO at different time. Under BSA-AGEO 400 mg / L, Different concentrations of simvastatin (5 渭 mol / L, 50 渭 mol / L) were exposed for 48 h, and BSA-AGEN 400 mg / L and simvastatin 50 渭 mol / L for 48 h. The supernatant was extracted and gelatinase method was used to determine the activity of MMP-9. The expression of MMP-9 was detected by Western blot. Statistical analysis.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R364.1

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